Tang Yiwei, Gao Jingwen, Liu Xiuying, Gao Xue, Ma Tao, Lu Xiaonan, Li Jianrong
College of Food Science & Project Engineering, Bohai University, Jinzhou 121013, China; Food, Nutrition, and Health Program, Faculty of Land and Food Systems, The University of British Columbia, Vancouver V6T 1Z4, Canada.
College of Food Science & Project Engineering, Bohai University, Jinzhou 121013, China.
Food Chem. 2017 Aug 1;228:62-69. doi: 10.1016/j.foodchem.2017.01.102. Epub 2017 Jan 20.
As an ideal biomimetic antibody, molecularly imprinted polymer (MIP) has shown great promise in immunoassays. Here, we developed a covalent imprinting method to prepare MIP artificial antibody towards clenbuterol on the well surface of a microtiter plate. Template molecule (clenbuterol) was splited by hydrolysis with functional monomer and removed by methanol/acetic acid solution, and then three-dimensional cavities were fabricated in the MIP, which can rebind template molecule via hydrogen bond interaction. Using this artificial antibody, we developed a novel biomimetic enzyme-linked immunosorbent assay (ELISA) with excellent sensitivity and specificity. The linear range for clenbuterol was 10 to 1000μgL, with a detection limit of 10μgL. Cross reactivity of this MIP artificial antibody to other four structural analogs was less than 0.93%. This method was applied to determine clenbuterol in real samples with satisfactory result, suggesting a promising application of the biomimetic ELISA in food and environmental analysis.
作为一种理想的仿生抗体，分子印迹聚合物（MIP）在免疫分析中显示出巨大的潜力。在此，我们开发了一种共价印迹方法，在微量滴定板的孔表面制备针对克伦特罗的MIP人工抗体。模板分子（克伦特罗）通过与功能单体水解而裂解，并用甲醇/乙酸溶液去除，然后在MIP中形成三维空腔，其可通过氢键相互作用重新结合模板分子。使用这种人工抗体，我们开发了一种新型的具有优异灵敏度和特异性的仿生酶联免疫吸附测定（ELISA）。克伦特罗的线性范围为10至1000μg/L，检测限为10μg/L。这种MIP人工抗体对其他四种结构类似物的交叉反应性小于0.93%。该方法应用于实际样品中克伦特罗的测定，结果令人满意，表明仿生ELISA在食品和环境分析中具有广阔的应用前景。
